Bradykinin antagonist peptides

ABSTRACT

The substitution of the L-Pro at the 7-position of the peptide hormone bradykinin or other substituted analogs of bradykinin with an aromatic amino acid or the D-configuration converts bradykinin agonists into a bradykinin antagonist. The invention further includes additional modifications at other positions within the novel 7-position modified bradykinin antagonists which increase enzyme resistance, antagonist potency and/or specificity of the new bradykinin antagonists. The analogs produced are useful in treating conditions and diseases of the mammal and human body in which an excess of bradykinin or related kinins are produced or injected as by bites into the body.

FIELD OF THE INVENTION

The invention relates to novel biologically active peptides which act asantagonists of the biological activities of bradykinin, theirpharmaceutically acceptable salts, and their application as therapeuticagents.

BACKGROUND OF THE INVENTION

In the 25 years since the sequence of the potent mammalian vasodilatorpeptide bradykinin was described and synthesized (Boissonnas et al.,Experientia 16: 326, 1960) several hundred sequence-related peptideanalogs have been synthesized and assayed in biological systems(Schroeder, in Handbook of Experimental Pharmacology, Vol. 25, (SpringerVerlag) pp 324-350, 1970) (Stewart, Handbook of ExperimentalPharmacology, Vol. 25 (Supplement), (Springer Verlag) pp 227-272, 1979).The objective in these studies was to investigate the variedphysiological and pharmacological roles of bradykinin.

Bradykinin, and its physiologically important related peptides kallidin(Lys-bradykinin) and Met-Lys-bradykinin, contract smooth muscle, (forexample to produce diarrhea and inflammatory bowel disease and asthma)lower blood pressure, mediate inflammation as in allergies, arthritisand asthma, participate in blood-clotting and complement-mediatedreactions in the body, mediate rhinitis (viral, allergic andnon-allergic) and are overproduced, in pathological conditions such asseptic shock, acute pancreatitis, hereditary angioneurotic edema,post-gastrectomy dumping syndrome, carcinoid syndrome, anaphylacticshock, reduced sperm motility, and certain other conditions. Theproduction of bradykinin from the plasma results in pain at the site ofthe pathological condition, and the overproduction intensifies the paindirectly or via stimulation by bradykinin of the activation of thearachidonic acid pathway which produces prostaglandins and leukotrienes,the more distal and actual mediators of inflammation. Literaturereferences describing these actions of bradykinin and related peptidesare found in Handbook of Experimental Pharmacology, Vol. 25,Springer-Verlag, 1970 and Vol. 25 Supplement, 1979.

Bradykinin as discussed has been found to be produced in inflammatoryreactions in the intestine provoking contraction of smooth muscle andsecretion of fluid and ions. The existence of specific bradykininreceptors in the mucosal lining of the intestine and intestinal smoothmuscle is demonstrated by Manning, et al. in Nature (229:256-259, 1982)showing the influence of bradykinin in very low concentrations uponfluid and ion secretion.

The production of bradykinin and associated pain in angina has beenstudied and reported by Kimura, et al. in American Heart Journal(85:635-647, 1973) and by Staszewska - Barczak, et al. in CardiovascularResearch (10:314-327, 1976). The reported action of bradykinin andprostaglandins acting in concert are the natural stimulus for excitationof the sensory receptors signalling the pain of myocardial ischeamia.

Bradykinin and bradykinin - related kinins are not only produced by theanimal but may also be injected as a result of stings and bites. It isknown that insects such as hornets and wasps inject bradykinin relatedpeptides which also cause pain, swelling and inflammation.

The search for understanding of the mechanism of action of bradykinin,which is essential for the development of useful tools for diagnosticuse, and for the development of therapeutic agents aimed at alleviatingthe intense pain caused by the production and overproduction ofbradykinin, has been severely hindered by the lack of specificsequence-related competitive antagonists of bradykinin.

Several non-peptide, non-specific and non-selective antagonists of oneor more of the biological activities of bradykinin have been describedamong compounds as diverse as analgesics and anti-inflammatorysubstances, which act via the prostaglandin system and not directly onbradykinin biological receptors (Rocha e Silva and Leme, Med. Exp. 8:287, 1963). These are antihistamines (Gecse et al., J. Pharm. Pharmacol.21: 544, 1969); bradykinin-antibodies (Grez et al., Eu. J. Pharmacol.29: 35, 1974); benzodiazepine derivatives (Leme and Rocha e Silva, Br.J. Pharmacol. 25: 50, 1965); high molecular weight ethylene oxidepolymers (Wilkens and Back, Arch. Intl. Pharmacodynam. 209: 305, 1974);gallic acid esters (Posati et al., J. Agri. Food Chem. 18: 632, 1970)and serotonin inhibitors (Gomazkon and Shimkovich, Bull. Exptl. Biol.Med. 80: 6, 1975). None of these individual compounds or classes ofcompounds specifically inhibit bradykinin.

Heptyl esters of various amino acid-containing substances, such assingle basic amino acids (i.e. Arg and Lys) (Gecse, Adv. Exptl. Biol.Med. 70: 5, 1976), the dipeptide Phe-Gly (Gecse et al., Int Aech.Allergy 41: 174, 1971), and of analogs of C- terminal peptide fragmentsof bradykinin (i.e. Pro-Phe-Arg)(Claesson et al., Adv. Exptl. Med. Biol.120B: 691, 1979) have been reported as anti-bradykinin substances. Whentested in bradykinin assay systems they prove to be weak partialagonists/antagonists, depending on the dose, with little specificity forinhibiting bradykinin action.

Preparations of damaged vascular tissue have been reported to respond tobradykinin analogs which lack the C-terminal Arg residue, but not tobradykinin itself, and analogs of these des-Arg⁹ -bradykinins have beendeveloped as antagonists of this non-physiological activity ofbradykinin. These antagonists have no significant bradykinin-likeagonist effects, nor any antagonist effect on any of the physiologicallysignificant kinin-responding systems (Regoli and Barabe, Pharmacol.Revs. 32:1,1980).

Several bradykinin analogs containing the O-methyl ether of Tyr residuesat positions 5 and/or 8 have been reported to produce mixedagonist/antagonist activity on isolated uteri of galactosemic rats, butnot on normal rats. The antagonism was not reliably reproducible inthese animals (Stewart and Woolley, in Hypotensive peptides, SpringerVerlag, pp 23-33, 1966).

Other changes in the bradykinin molecule have been additions of aminoacids at the N-terminal end which affect the rate of enzymaticdegradation of bradykinin in vivo.

The half life of bradykinin in the systemic circulation is less than 30seconds (S. H. Ferreira & J. R. Vane, Br. J. Pharmacol. Chemotherap.30:417, 1967). Bradykinin is completely destroyed (98-99% destruction)on a single passage through the pulmonary circulation (J. Roblero, J. W.Ryan and J. M. Stewart, Res. Commun. Pathol. Pharmacol. 6:207, 1973) asdetermined in the anesthetized rat by measuring the depressor effects ofan agonist following intraaortic (IA) (bypassing the pulmonarycirculation) and intravenous (IV) administration. Resistance ofbradykinin agonists to pulmonary kininase destruction in vivo ispromoted by addition of single (ie, DArg-, DLys-, Lys-) and double(DLys-Lys-) basic amino acid residues to the N-terminal of thebradykinin sequence. The addition of the dipeptide Lys-Lys to theN-terminal of bradykinin agonists confers complete resistance to in vivodestruction on initial passage through the pulmonary circulation(Roblero, Ryan and Stewart, Res. Comm. Pathol. Pharmacol. 6:207, 1973).

SUMMARY OF INVENTION

The invention relates to the modification of the sequence of themammalian peptide hormone bradykinin(Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) and pharmaceutically acceptablesalts thereof, at the Pro residue at position 7 in a unique mannerwhich, for the first time, produces sequence-related analogues that actas specific and competitive inhibitors of the biological activities ofbradykinin. The invention specifically relates to the substitution ofthe L-Pro at position 7 with aromatic amino acids of theD-configuration, a change which converts bradykinin agonists intoantagonists, and includes additional modifications at other positionswithin the 7-position modified bradykinin antagonist which conferincreased antagonist potency, resistance to enzymatic degradation and/ortissue specificity on the D-amino acid-containing bradykinin sequence.More specifically, the invention relates to the peptides of the generalformula: ##STR1## Wherein A is a hydrogen atom or single acidic, basic,neutral or aromatic amino acid residue of the D-or L-configuration, suchas D-Arg, D-Lys or L-Thi, an N-terminal enzyme protecting group from thegroup comprising acyl-type protecting groups, aromatic urethane-typeprotecting groups, alkyl-type protecting groups, or alternately A is adi- or poly-peptide containing amino acids of the D- or L-configuration, such as Lys-Lys, Met-Lys, or Gly-Arg-Met-Lys;

B is an L-pro residue, or other D- or L-cyclic or noncyclic aliphaticamino acid residue, such as L-hydroxyproline, an L-aromatic orsubstituted aromatic amino acid residue;

C is D- or L-Pro residue, or other cyclic, aliphatic, aromatic orsubstituted aromatic amino acid residue of the D- or L-configuration;

D is a Gly residue or other aliphatic, aromatic or substituted aromaticamino acid residue of the L-configuration, such as Ala;

W is a Phe residue of the L-configuration, or a substituted Phe or otheraliphatic or aromatic amino acid residue, such as Leu,beta-2-thienyl-alanine (Thi) or 2-pyridylalanine (Pal);

X is a Ser residue of the L-configuration, a Gly residue, or other D- orL-aliphatic or aromatic or substituted aromatic amino acid residue, suchas pCl-D-Phe or D-Phe;

Y is a D-aromatic amino acid residue, or substituted aromatic amino acidresidue, such as D-Phe, beta-(2-thienyl)-D Ala (DThi),beta-(2-pyridyl)-D-Ala (D-Pal), β-2-naphthyl-D Ala (D-Nal), DHis,D-homo-Phe (DhPhe), O-methyl-DTyr (DOMT), D-alphaphenyl-Gly (DPhg),DTrp, DTyr or pCl-DPhe (CDF);

Z is a Phe residue of the L-configuration, or a substituted Phe or otheraliphatic or aromatic amino acid residue, such as Leu, Thi or Pal.

In a preferred compound of the general formula I the substituents havethe following identity: A=H, B=Pro or Hyp, C=Pro or Hyp, D=Gly, W=Z=Pheor Thi, X=Ser and Y=any aromatic amino acid of the D- configuration.

Salts of peptides of general formula I include salts with HCl, TFA,AcOH, as well as other pharmaceutically acceptable salts.

The following TABLES I and II show substitutions that can be made in thebradykinin polypeptide and the effect of such substitutions. Indicatedsubstitutions of the 0, 1, 2, 3, 5, 7 and 8 amino acid residues ofbradykinin yield preferred bradykinin antagonists.

                  TABLE I                                                         ______________________________________                                        SUBSTITUTIONS IN BRADYDININ ANTAGONISTS                                       ______________________________________                                         ##STR2##                                                                     ______________________________________                                         Thi = β-(2Thienyl)alanine?                                               Pal = β-(2Pyridyl)alanine?                                               Hyp = 4Hydroxyproline                                                         Azt = Azetidine2-carboxylic acid                                              Thz = Thiazolidine2-carboxylic acid                                           Inip = Isonipecotic acid                                                      OMT = OMethyltyrosine                                                         CDF = parachloro-D-phenylalanine                                              Nal = β-(2Naphthyl)-alanine                                              CLF = parachloro-L-phenylalanine                                              PNF = paranitrophenylalanine                                                  ΔPro = 2, 3Dehydroproline                                          

                  TABLE II                                                        ______________________________________                                        CHARACTERISTICS OF BRADYKININ ANTAGONISTS                                     ______________________________________                                         ##STR3##                                                                 

DETAILED DESCRIPTION

The synthesis of the peptides of general Formula I, includingderivatization, activation, and coupling of protected amino acidresidues, and their purification, and the analytical methods fordetermining identity and purity are included in the general body ofknowledge of peptide chemistry, as described in Houben Weyl "Methodender Organische Chemie" Vol. 16, parts I & II(1974) for solution-phasesynthesis, and in "Solid Phase Peptide Synthesis" by Stewart and Young(1984) for synthesis by the solid-phase method of Merrifield. Anychemist skilled in the art of peptide synthesis can synthesize thepeptides of general Formula I by standard solution methods or by manualor automated solid-phase methods.

The symbols and abbreviations used for amino acids, their derivativesand protecting groups, and peptides and their salts are thosecustomarily used in peptide chemistry (Biochem. J. 126:773,1972, theJournal reference is hereby incorporated by reference.). For convenienceseveral abbreviations are defined in Table III reproduced below. Allamino acid residues, except Gly, described in the specification but notthe claims are of the L-configuration unless otherwise specified.

                  TABLE III                                                       ______________________________________                                        ABBREVIATIONS FOR AMINO ACID RESIDUES                                         ______________________________________                                        Aib            alpha-aminoisobutyric acid                                     Azt            azetidine-2-carboxylic acid                                    CDF            para-chloro-D-phenylalanine                                    CLF            para-chloro-L-phenylalanine                                    hPhe           homo-phenylalanine                                             Hyp            4-hydroxy-proline                                              Inip           isonipecotic acid                                              MDY            O--methyl-D-tyrosine                                           Nal            beta-(2-naphthyl)-alanine                                      ΔPro     2,3-dehydroproline                                             Pal            beta-(3-pyridyl)-alanine                                       Phg            alpha-phenylglycine                                            Sar            sarcosine                                                      Thi            beta-(2-thienyl)-alanine                                       Thz            thiazolidine-2-carboxylic acid                                 ______________________________________                                         (all other abbreviations follow the IUPAC standards for amino acid            residues)                                                                

The following examples are illustrative of compounds of this inventionwith general formula I and are not limitative. All percentages andratios are by weight when solids are involved and by volume when onlyliquids are involved.

EXAMPLE 1

Preparation of Arg-Pro-Pro-Gly-Phe-Ser-DPhe-Phe-Arg (DPhe -BK).

A mixture of 6.4 gm of tertiary butyloxy carbonyl- (g-paratoluenesulfonyl)-Arg [BOC-Arg(Tos)](15 mMole) and 183 mg of N,N-dimethylaminopyridine (1.5 mMole) was dissolved in a mixture of 20 mlof dimethylformamide (DMF) and 125 ml of dichloromethane (DCM). Fifteeng of hydroxymethyl-polystyrene-divinyl benzene (1% crosslinked,containing 0.74 mMole of free hydroxyl group per g of resin) was added,followed by 60 ml of a 0.25 M solution of dicyclohexylcarbodiimide (DCC)in DCM at room temperature. The suspension was stirred at roomtemperature overnight, filtered, and the resin was washed three timeswith 60 ml of DCM, three times with 60 ml of methyl alcohol (MeOH), andreswollen in 120 ml of DCM. The coupling of another portion ofBOC-Arg(Tos) was conducted on the resin as above. After filtering andwashing the resin it was reswollen in 120 ml of DCM, and 2.1 ml ofbenzoyl chloride and 1.5 ml of triethylamine (Et₃ N) were added. Afterstirring the suspension for 30 minutes at room temperature the resin wasfiltered, washed three times with 60 ml portions of DCM, MeOH, washedthree times with 60 ml portion of MeOH and finally washed three timeswith 60 ml portions of DCM. The resin was air dried to constant weightto give 18.5 gm of BOC-Arg(Tos)-hydroxymethyl-resin, with an actualamino acid content of 0.272 millimoles of Arg per g of resin asdetermined by quantitative amino acid analysis of a sample of the aminoacid resin following hydrolysis (4 hr, 130° C.) in 6N HCL/propionicacid.

The resin, 1.5 gm containing a total of 0.4 mMole of Arg, was placed inthe reaction vessel of an automatic solidphase synthesizer (Beckmanmodel 990) and subjected to one cycle of addition for the coupling ofBOC-Phe as follows;

PROGRAM A. STANDARD DCC COUPLING:

The resin was washed three times with 20 ml portions of DCM. The resinwas then equilibrated with 20 ml of a 1:3 ratio of trifluoroacetic acid(TFA) in DCM containing 0.1% indole for 1.5 minutes. The equilibrationwas then repeated for 30 minutes. The resin was then washed six timeswith 20 ml portions of DCM followed by neutralization with a 10%solution of (Et₃ N) in DCM for one and one half minutes, then theneutralization step was repeated. The resin was washed six times with 20ml of DCM and then equilibrated with a solution of 1.0 mMole of BOC-Phein DCM for one and one half minutes. Then four ml of 0.25N DCC in DCMwas added and the mixture stirred for two hours. Then the resin waswashed three times with 20 ml portions of DCM.

A second cycle of addition was performed according to Program B:

REVERSE ADDITION:

The procedure of Program A through neutralization and following washeswas repeated. Then 1.0 mMole of DCC in 4 ml of DCM was added and theresin and solution were mixed for one and one-half minutes. Then 1.0mMole of BOC-D-Phe in 12 ml DCM was added and the resin and solutionwere mixed for two hours. The resin was then washed six times with 20 mlportions of DCM.

The N-Terminal protecting group was removed according to the followingsequence:

PROGRAM C. TERMINAL DEPROTECTION:

The procedure of PROGRAM A up to the neutralization with triethylaminewas repeated. The resin was then washed 6 times with 20 ml portions ofethyl alcohol and the peptide-resin was air dried giving 1.66 gm ofDPhe-Phe-Arg-Resin as the trifluoroacetic acid salt.

Synthesis was continued with 410 mg of the DPhe-Phe-Arg-Resin TFA salt.The next residue was added according to PROGRAM D.

PROGRAM D. RECOUPLE:

The peptide-resin salt was first washed three times with 20 ml portionof DCM, then neutralized with 10% Et₃ N DCM for 1.5 minutes. Theneutralization step was then repeated and the peptide-resin-salt waswashed six times with 20 ml portions of DCM. The peptide-resin was thenequilibrated with a solution of 1.0 mMole of BOC-Ser(0 Bzl) in DMF for1.5 minutes. Four ml of 0.25N DCC in DCM was added and mixed with theresin for two hours. The product was washed three times with DCM.

The following amino acid derivatives were added to the growing peptidechain according to the listed Programs: BOC-Phe (A), BOC-Gly (A),BOC-Pro (A), BOC-Pro (A), followed by recouple of BOC-Pro (D),BOC-Arg(Tos)(dissolved in 2 ml DMF +9 ml DCM), (A), followed by ProgramC. This gave 530 mg of protected nonapeptide-resin as the TFA salt.

A 510 mg portion of the peptide-resin above was suspended in 10 ml ofliquid anhydrous HF containing 1 ml of anisole at -70° C. and stirred 45min. at 0° C. HF and anisole were removed by vacuum (1 hr water pump, 1hr vacuum pump), the peptide plus resin was washed three times with 20ml portions of ethyl ether (Et₂ O) and the peptide extracted intoglacial acetic acid using three 6 ml extractions. The acetic acidsolution was lyophilized to give 185 mg of crude deprotected peptide.

The peptide was purified by countercurrent distribution (CCD) (100 upperphase transfers in a Post CCD apparatus) in the solvent system nBuOH:1%TFA (1:1). The content of the tubes corresponding to the mainpeptide-containing peak, as determined by the quantitative Sakaguchireagent, was collected, the solvent evaporated under reduced pressure,the residue dissolved in glacial acetic acid (AcOH) and lyophilized togive 140 mg of peptide with a partition coefficient (k) from the CCD of5.7. Repeating the countercurrent distribution in the solvent systemnBuOH:AcOH:H2O(4:1:5) gave, upon detection and workup as describedabove, 73 mg of Arg-Pro-Pro-Gly-Phe-Ser-DPhe-Phe-Arg as the TFA salt(k=0.2). Thin layer chromatographs (TLC) on Merck glass precoated silicagel plates in the solvent systems nBuOH:AcOH:H2O(8:3:4) andEtOAc:pyridine:AcOH:H2O(5:5:1:3) gave Rf(834) of 0.17 and Rf(5513) of0.36 for the pure peptide, as visualized by the chlorine-tolidinepeptide identification spray. Quantitative amino acid analysis (Beckman120 instrument) after acid hydrolysis (17 hr in sealed glass vials underN₂ at 110° C. in 2 ml 6N HCl containing 2 drops 2-mercaptoethanol and 40microliters of phenol) gave the following ratios of amino acids:Arg(2.12); Pro(1.93); Gly(1.01); Phe(2.98); Ser(0.96).

EXAMPLE 2

Preparation of Arg-Pro-Pro-Gly-Phe-Ser-DThi-Phe-Arg (DThi -BK).

This peptide was prepared by the method in Example 1, except thatBOC-beta-2-thienyl-D-Ala (BOC-DThi) was used in place of BOC-DPhe:k(415)=0.24; Arg(2.02), Pro(2.18), Gly(1.00), Phe(1.99), Ser(0.89),Thi(0.99).

EXAMPLE 3

Preparation of Arg-Pro-Pro-Gly-Phe-Ser-DPal-Phe-Arg (DPal -BK).

This peptide was prepared by the method in Example 1, except thatBOC-2-pyridyl-DAla (BOC-Dpal) was used in place of BOC-DPhe:k(1:1)=0.22; Arg(2.03), Pro(2.01), Gly(1.02), Phe(1.99), Pal(1.02).

EXAMPLE 4

Preparation of Arg-Pro-Pro-Gly-Phe-DPhe-DPhe-Phe-Arg (DPhe -BK).

This peptide was prepared by the method in Example 1, except thatBOC-DPhe was used in place of BOC-Ser(Bzl): k(415)=1.3; Arg(2.12),Pro(1.94), Gly(1.00), Phe(3.94).

EXAMPLE 5

Preparation of Arg-Pro-Pro-Gly-Phe-DPhe-CDF-Phe-Arg (DPhe⁶ CDF⁷ -BK).

This peptide was prepared by the method in Example 1, except thatBOC-DPhe was used in place of BOC-Ser(Bzl) and BOC-pCl-D-Phe (BOC-CDF)was used in place of BOC-DPhe: k(415)=0.82; Arg(1.98), Pro(1.93),Gly(0.97), Phe(3.10), CDF(1.02).

EXAMPLE 6

Preparation of Arg-Pro-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Thi⁵,8 DPhe⁷ -BK).

This peptide was prepared by the method in Example 1, except thatBOC-beta-2-thienyl-Ala (BOC-Thi) was used in the two addition cycles inwhich BOC-Phe was used in Example 1: k(415)=0.21; Arg(1.98), Pro(1.94),Gly(1.04), Phe(1.04), Ser(0.96), Thi(2.05).

EXAMPLE 7

Preparation of Arg-Pro-Pro-Gly-Thi-Ser-DThi-Thi-Arg (Thi⁵,8 DThi⁷ -BK).

This peptide was prepared by the method in Example 6, except thatBOC-DThi was used in place of BOC-DPhe: k(415)=018; Arg(2.07),Pro(2.08), Gly(1.00), Ser(0.93), Thi(2.88).

EXAMPLE 8

Preparation of Arg-Pro-Pro-Gly-Thi-Ser-DPal-Thi-Arg (Thi⁵,8 DPal⁷ -BK).

This peptide was prepared by the method in Example 6, except thatBOC-DPal was used in place of BOC-DPhe: k(1:1)=0.15; Arg(2.00),Pro(2.20), Gly(1.09), Ser(0.89), Thi(1.92), Pal(0.89).

EXAMPLE 9

Preparation of Arg-Pro-Pro-Gly-Thi-DPhe-CDF-Thi-Arg (Thi⁵,8 DPhe⁶ CDF⁷-BK).

This peptide was prepared by the method in Example 5, except thatBOC-Thi was used in two cycles of addition in place of BOC-Phe:k(415)=0.75; Arg(1.89), Pro(2.08), Gly(1.06), Phe(1.02), Thi(1.88),CDF(1.07).

EXAMPLE 10

Preparation of DArg-Arg-Pro-Pro-Gly-Phe-Ser-DPhe-Phe-Arg (DArg⁰ DPhe⁷-BK).

This peptide was prepared by the method in Example 1, except that oneadditional cycle using BOC-(Tos)-DArg was performed with Program Afollowed by terminal deprotection with Program C: k(1:1)=3.55;Arg(2.89), Pro(2.07), Gly(1.02), Phe(3.05), Ser(0.98).

EXAMPLE 11

Preparation of DArg-Arg-Pro-DPro-Gly-Phe-Ser-DPhe-Phe-Arg (DArg⁰ Pro³DPhe⁷ -BK).

This peptide was prepared by the method in Example 10, except thatBOC-DPro was used in place of BOC-Pro in the first addition of BOC-Pro:k(415)=0.15; Arg(3.12), Pro(1.90), Gly(1.05), Phe(3.02), Ser(0.92).

EXAMPLE 12

Preparation of Arg-Pro-DPro-Gly-Thi-DPhe-CDF-Thi-Arg (DPro³ Thi⁵,8 DPhe⁶CDF⁷ -BK).

This peptide was prepared by the method in Example 9, except thatBOC-DPro was used in place of BOC-Pro in the first addition of BOC-Pro:k(415)=0.18; Arg(2.00), Pro(1.98), Gly(1.04), Phe(0.99), Thi(1.89),PCF(1.11).

EXAMPLE 13

Preparation of Lys-Lys-Arg-Pro-Pro-Gly-Phe-Ser-DPhe-Phe-Arg(Lys-Lys-DPhe⁷ -BK).

This peptide was prepared by the method in Example 1 except that twoadditional cycles of addition were performed with BOC-(e-ClZ)Lys, thefirst with Program D, the second with Program A followed by Program C:k(1:1)=0.52; Arg(2.04), Pro(1.99), Gly(0.96), Phe(3.00), Ser(0.96),Lys(2.01).

EXAMPLE 14

Preparation of Lys-Lys-Arg-Pro-Pro-Gly-Thi-Ser-DPhe-Thi-Arg(Lys-Lys-Thi⁵,8 DPhe⁷ -BK).

This peptide was prepared by the method in Example 13, except thatBOC-Thi was used in place of BOC-Phe in the two cycles of addition ofBOC-Phe: k(1:1)=0.33; Arg(1.98), Pro(1.97), Gly(1.01), Phe(1.03),Ser(0.97), Thi(1.96), Lys(2.08).

EXAMPLE 15

Preparation of DArg-Arg-Pro-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (DArg⁰ Thi⁵,8DPhe⁷ -BK).

This peptide was produced by the method in Example 6, except that oneadditional cycle of addition with BOC-(Tos)DArg was performed withProgram D, followed by Program C: k(1:1)=2.33; Arg(3.00), Pro(1.99),Gly(0.96), Phe(0.99), Ser(0.95),

EXAMPLE 16

Preparation of DArg-Arg-Pro-DPro-Gly-Thi-Ser-DPhe-Thi-Arg (DArg⁰ DPro⁵,8DPhe⁷ -BK).

This peptide was produced by the method in Example 15, except thatBOC-DPro was used in place of BOC-Pro in the first addition of BOC-Pro:k(415)=0.22; Arg(2.10), Pro(1.96), Gly(1.05), Phe(0.98), Ser(0.94),Thi(1.96).

EXAMPLE 17

Preparation of Lys-Lys-Arg-Pro-Hyp-Gly-Phe-Ser-DPhe-Phe-Arg(Lys-Lys-Hyp³ DPhe⁷ -BK).

This peptide was prepared by the method in Example 13, except thatBOC-(4-hydroxy)-Pro (BOC-Hyp) was used in place of BOC-Pro in the firstaddition of BOC-Pro: k(1:1)=0.35; Arg(2.00), Pro(1.03), Gly(1.00),Phe(3.08), Ser(0.94), Lys(1.95), Hyp(1.01).

EXAMPLE 18

Preparation of Arg-Hyp-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Hyp² Thi⁵,8 DPhe⁷-BK).

This peptide was produced by the method in Example 6, except thatBOC-Hyp was used in place of BOC-Pro in the second addition of BOC-Pro,including the recoupling with Program D: k(1:1)=2.45; Arg(2.03),Pro(0.98), Gly(1.06), Phe(1.05), Ser(0.95), Thi(1.97), Hyp(0.95).

EXAMPLE 19

Preparation of Lys-Lys-Arg-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Lys-Lys-Hyp²Thi⁵,8 DPhe⁷ -BK).

This peptide was produced by the method in Example 18, except that twoadditional cycles of addition with BOC-(e-ClZ)Lys were performed, thefirst with Program D, the second with Program A followed by Program C:k(1:1)=0.27; Arg(2.07), Pro(0.92), Gly(0.99), Phe(1.03), Ser(0.93),Thi(1.92), Lys(2.06), Hyp(1.10).

EXAMPLE 20

Preparation of Arg-Pro-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Hyp³ Thi⁵,8 DPhe⁷-BK).

This peptide was produced by the method in Example 6, except thatBOC-Hyp was used in place of BOC-Pro in the first addition of BOC-Pro:k(1:1)=2.23; Arg(2.08), Pro(0.98), Gly(1.04), Phe(1.01), Ser(0.99),Thi(1.95), Hyp(0.94).

EXAMPLE 21

Preparation of DArg-Arg-Pro-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (DArg⁰ -Hyp³Thi⁵,8 DPhe⁷ -BK).

This peptide was produced by the method in Example 20, except that oneadditional cycle of addition with BOC-(Tos)DArg was done with Program D,followed by Program C: k(1:1)=0.18; Arg (3.01), Pro(1.02), Gly(0.98),Phe(1.02), Ser(0.92), Thi(2.07), Hyp(0.97).

EXAMPLE 22

Preparation of Arg-Hyp-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Hyp²,3 Thi⁵,8 DPhe⁷-BK).

This peptide was produced by the method in Example 6, except thatBOC-Hyp was used in place of BOC-Pro in those cycles where BOC-Pro hadbeen used: k(1:1)=1.56; Arg(2.04), Gly(1.06), Phe(1.02), Ser(1.01),Thi(1.94), Hyp(1.93).

EXAMPLE 23

Preparation of Arg-Hyp-Hyp-Gly-Thi-DPhe-CDF-Thi-Arg (Hyp²,3 Thi⁵,8 DPhe⁶CDF⁷ -BK).

This peptide was produced by the method in Example 9, except thatBOC-Hyp was used in place of BOC-Pro in those cycles where BOC-Pro hadbeen used: k(415)=0.89; Arg(2.06), Gly(1.00), Phe(1.03), PCF(1.05),Thi(1.92), Hyp(1.93).

EXAMPLE 24

Preparation of Arg-Pro-Pro-Gly-Leu-Gly-DPhe-Leu-Arg (Gly⁶ Leu⁵,8 DPhe⁷-BK).

This peptide was produced by the method in Example 1, except thatBOC-Leu was used in place of BOC-Phe in those cycles where BOC-Phe hadbeen used, and BOC-Gly was used when BOC-(OBzl)Ser had been used:k(415)=0.30; Arg(2.04), Pro(2.05), Gly(1.98), Phe(0.98), Leu(1.95).

EXAMPLES 25-103

The following examples were prepared by methods identical to the methodsdescribed above for similarly substituted peptides, and are notlimitative:

25. Arg-Pro-Pro-Gly-Phe-Ser-DNal-Phe-Arg (DNal⁷ -BK): k(415)=0.37;Arg(2.09), Pro(2.02), Gly(0.98), Phe(2.06), Ser(0.96), Nal(0.95).

26. Arg-Pro-Pro-Gly-Phe-Ser-MDY-Phe-Arg (MDY⁷ -BK): k(1:1)=4.88;Arg(2.10), Pro(1.91), Gly(0.96), Phe(2.08), Ser(0.94), MDY(1.04).

27. Arg-Pro-Pro-Gly-Phe-Ser-DPhg-Phe-Arg (DPhg⁷ -BK): k(1:1)=3.55;Arg(2.01), Pro(1.90), Gly(1.03), Phe(2.07), Ser(0.99),

28. Arg-Pro-Pro-Gly-Phe-Ser-DHis-Phe-Arg (DHis⁷ -BK): k(1:1)=0.30;Arg(2.04), Pro(2.09), Gly(0.94), Phe(2.00), Ser(1.00), His(0.93).

29. Arg-Pro-Pro-Gly-Phe-Ser-DTrp-Phe-Arg (DTrp⁷ -BK): k(415)=0.30;Arg(2.04), Pro(1.95), Gly(1.02), Phe(2.05), Ser(0.95), TrP(0.98).

30. Arg-Pro-Pro-Gly-Phe-Ser-DTyr-Phe-Arg (DTyr⁷ -BK): k(1:1)=2.70;Arg(1.94), Pro(1.88), Gly(1.04), Phe(2.10), Ser(0.97), Tyr(1.08).

31. Arg-Pro-Pro-Gly-Phe-Ser-DhPhe-Phe-Arg (DhPhe⁷ -BK): k(415)=0.37;Arg(1.99), Pro(1.94), Gly(0.97), Phe(2.02), Ser(0.88), hPhe(1.20).

32. Arg-Pro-Pro-Gly-Phe-DPhe-DThi-Phe-Arg (DPhe⁶ DThi⁷ -BK):k(415)=0.59; Arg(2.14), Pro(1.87), Gly(1.00), Phe(3.01), Thi(0.98).

33. Arg-Pro-Pro-Gly-Phe-DThi-DThi-Phe-Arg (DThi⁶,7 -BK): k(415)= 0.54;Arg(2.04), Pro(2.00), Gly(1.01), Phe(2.04), Thi(1.91).

34. Arg-Pro-Pro-Gly-Phe-DPhe-DNal-Phe-Arg (DPhe⁶ DNal⁷ -BK): k(415)=1.04; Arg(2.02), Pro(2.04), Gly(1.01), Phe(2.94), Nal(0.99).

35. Arg-Pro-Pro-Gly-Phe-DPhe-MDY-Phe-Arg (DPhe⁶ MDY⁷ -BK): k(415)=0.59;Arg(2.11), Pro(1.90), Gly(0.97), Phe(3.02), MDY(1.01).

36. Arg-Pro-Pro-Gly-Phe-DPhe-DPal-Phe-Arg (DPhe⁶ DPal⁷ -BK): k(415)=0.16; Arg(1.86), Pro(2.12), Gly(1.07), Phe(2.90), Pal(1.05).

37. Arg-Pro-Pro-Gly-Phe-Gly-DVal-Phe-Arg (Gly⁶ DVal⁷ -BK): k(415)=0.20;Arg(2.08), Pro(2.00), Gly(1.96), Phe(1.98), Val (0.98).

38. Arg-Hyp-Pro-Gly-Phe-Ser-DPhe-Phe-Arg (Hyp² DPhe⁷ -BK): k(1:1)=3.76;Arg(2.00), Pro(0.93), Gly(1.02), Phe(3.16), Ser(0.94), Hyp(0.94).

39. Arg-Pro-Hyp-Gly-Phe-Ser-DPhe-Phe-Arg (Hyp³ DPhe⁷ -BK): k(1:1)=3.35;Arg(1.96), Pro(0.97), Gly(1.01), Phe(3.10), Ser(0.94), Hyp(1.02). Hyp(

40 Arg-Hyp-Hyp-Gly-Phe-Ser-DPhe-Phe-Arg (Hyp²,3 DPhe⁷ -BK): k(1:1)=2.33;Arg(2.03), Gly(1.02), Phe(3.10), Ser(0.94), Hyp(1.91).

41. Arg-Pro-Pro-Gly-Thi-DPhe-DPhe-Thi-Arg (Thi⁵,8 DPhe⁶,7 -BK).

42. Arg-Pro-Pro-Gly-Thi-DThi-DThi-Thi-Arg (Thi⁵,8 DThi⁶,7 -BK):k(415)=0.37; Arg(2.08), Pro(2.08), Gly(0.99), Thi(3.86).

43. Arg-Pro-Pro-Gly-Thi-DThi-DPal-Thi-Arg (Thi⁵,8 DThi⁶ DPal⁷ -BK):k(1:1)=0.70; Arg(1.93), Pro(2.11), Gly(1.08), Thi(2.86), Pal(1.02).

44. Arg-Pro-Pro-Gly-Thi-DThi-DNal-Thi-Arg (Thi⁵,8 DThi⁶ DNal⁷ -BK):k(415)=0.59; Arg(2.19), Pro(2.08), Gly(1.00), Nal(0.93), Thi(2.88).

45. Arg-Pro-Pro-Gly-Thi-DThi-CDF-Thi-Arg (Thi⁵,8 DThi⁶ CDF⁷ -BK):k(415)=0.54; Arg(2.16), Pro(1.97), Gly(1.00), PCF(1.06), Thi(2.81).

46. Ar g-Pro-Pro-Gly-Thi-DPhe-DAla-Thi-Arg (Thi⁵,8 DPhe⁶ DAla⁷ -BK):k(415)=0.32; Arg(2.06), Pro(1.89), Gly(0.97), Phe(1.02), Ala(0.96),Thi(2.12).

47. Arg-Pro-Pro-Gly-Thi-CDF-DAla-Thi-Arg (Thi⁵,8 CDF⁶ DAla⁷ -BK):k(415)=0.39; Arg(1.96), Pro(1.93), Gly(1.00), Ala(0.99), Thi(2.05),PCF(1.08).

48. Thi-Arg-Pro-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Thi⁰ -Thi⁵,8 -DPhe⁷ -BK):k(415)=0.27; Arg(2.16), Pro(1.89), Gly(1.03), Phe(1.04), Ser(0.93),Thi(2.95).

49. DArg-Arg-Pro-Pro-Gly-Phe-Ser-DPhg-Phe-Arg (DArg⁰ -DPhg⁷ -BK):k(1:1)=2.03; Arg(3.05), Pro(2.00), Gly(0.97), Phe(2.04), Ser(0.94),Phg(1.00).

50. DArg-Arg-Pro-Pro-Gly-Phe-Ser-DTrp-Phe-Arg (DArg⁰ -DTrp⁷ -BK):k(1:1)=4.88; Arg(3.07), Pro(1.99), Gly(1.00), Phe(2.04), Ser(0.89),Trp(0.97).

51. DArg-Arg-Pro-Pro-Gly-Phe-Ser-DTyr-Phe-Arg (DArg⁰ -DTyr⁷ -BK):k(1:1)=1.56; Arg(2.96), Pro(2.09), Gly(1.03), Phe(1.99), Ser(0.90),Tyr(1.04).

52. DArg-Arg-Pro-Pro-Gly-Phe-Ser-DHis-Phe-Arg (DArg⁰ -DHis⁷ -BK):k(1:1)=0.18; Arg(3.08), Pro(1.99), Gly(1.02), Phe(2.07), Ser(0.89),His(0.96).

53. DArg-Arg-Pro-Pro-Gly-Phe-Ser-DhPhe-Phe-Arg (DArg⁰ -DhPhe⁷ -BK):k(415)=0.18; Arg(3.06), Pro(1.98), Gly(0.95), Phe(2.05), Ser(0.86),DhPhe(1.11).

54. Lys-Lys-Arg-Pro-Pro-Gly-Phe-Ser-DPhe-Phe-Arg (Lys-Lys-DPhe⁷ -BK):k(1:1)=0.52; Arg(2.04), Pro(2.01), Gly(0.96, Phe(3.00), Ser(0.96).

55. Lys-Lys-Arg-Pro-Pro-Gly-Phe-Ser-DTrp-Phe-Arg (Lys-Lys-DTrp⁷ -BK):k(1:1)=0.67; Arg(2.01), Pro(2.00), Gly(1.01), Phe(2.03), Ser(0.91),Trp(0.88), Lys(2.03).

56. Lys-Lys-Arg-Pro-Pro-Gly-Phe-Ser-DTyr-Phe-Arg (Lys-Lys-DTyr⁷ -BK):k(1:1)=0.21; Arg(2.01), Pro(2.11), Gly(0.99), Phe(2.02), Ser(0.97),Tyr(1.02), Lys(1.90).

57. Lys-Lys-Arg-Pro-Pro-Gly-Phe-Ser-DHis-Phe-Arg (Lys-Lys-DHis⁷ -BK):k(1:1)=0.08; Arg(1.91), Pro(2.06), Gly(0.94), Phe(2.08), Ser(0.93),His(0.95), Lys(2.12).

58. Lys-Lys-Arg-Pro-Pro-Gly-Phe-Ser-DhPhe-Phe-Arg (Lys-Lys-DhPhe⁷ -BK):k(1:1)=1.13; Arg(1.99), Pro(1.93), Gly(0.98), Phe(2.06), Ser(0.92),DhPhe(1.11), Lys(2.01).

59. Arg-Pro-DPro-Gly-Phe-Ser-DPhe-Phe-Arg (DPro³ DPhe⁷ -BK):k(415)=0.27; Arg(2.07), Pro(1.97), Gly(0.98), Phe(3.02), Ser(0.95).

60. Arg-Pro-DPro-Gly-Phe-CDF-DAla-Phe-Arg (DPro³ CDF⁶ DAla⁷ -BK):k(415)=0.59; Arg(1.99), Pro(2.03), Gly(0.97), Phe(2.02), Ala(0.99),PCF(1.02).

61. Arg-Pro-DPro-Gly-Thi-Ser-DPhe-Thi-Arg (DPro³ Thi⁵,8 DPhe⁷ -BK):k(415)=0.22; Arg(2.10), Pro(1.96), Gly(1.05), Phe(0.98), Ser(0.94),Thi(1.96).

62. DArg-Arg-Pro-Pro-Gly-Thi-DPhe-DAla-Thi-Arg (DArg⁰ -Thi⁵,8 DPhe⁶DAla⁷ -BK): k(1:1)=4.00; Arg(2.89), Pro(1.95), Gly(1.03), Phe(0.98),Ala(1.01), Thi(2.14).

63. DArg-Arg-Pro-Pro-Gly-Thi-CDF-DAla-Thi-Arg (DArg⁰ -Thi⁵,8 CDF⁶ DAla⁷-BK): k(415)=0.19; Arg(3.00), Pro(1.99), Gly(1.00), Ala(0.96),Thi(2.03), PCF(1.02).

64. DArg-Arg-Hyp-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (DArg⁰ Hyp² Thi⁵,8 DPhe⁷-Bk): k(1:1)=1.50; Arg(3.11), Pro(0.97), Gly(1.03), Phe(1.04),Ser(0.95), Thi(1.88), Hyp(1.02)

65. DArg-Arg-Hyp-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (DArg⁰ -Hyp²,3 Thi⁵,8DPhe⁷ -Bk): k(1:1)=0.96; Arg(3.19), Gly(0.97), Phe(0.98), Ser(1.00),Thi(1.95), Hyp(1.90)

66. Thi-Arg-Hyp-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Thi⁰ -Hyp² Thi⁵,8 DPhe⁷-BK)

67. Thi-Arg-Pro-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Thi⁰ -Hyp³ Thi⁵,8 DPhe⁷-Bk)

68. Thi-Arg-Hyp-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Thi⁰ -Hyp²,3 Thi⁵,8 DPhe⁷-Bk)

69. Arg-Hyp-Pro-Gly-Thi-DPhe-CDF-Thi-Arg (Hyp² Thi⁵,8 DPhe⁶ CDF⁷ -BK):k(415)=1.04; Arg(2.02), Pro(1.01), Gly(1.00), Phe(1.01), PCF(1.09),Thi(1.89), Hyp(0.98)

70. Arg-Pro-Hyp-Gly-Thi-DPhe-CFD-Thi-Arg (Hyp³ Thi⁵,8 DPhe⁶ CDF⁷ -BK):k(1:1)=0.96; Arg(2.01), Pro(1.11), Gly(0.97), Phe(1.06), Thi(1.86),PCF(1.06), Hyp(0.95)

71. Arg-Hyp-DPro-Gly-Thi-DPhe-CDF-Thi-Arg (Hyp² Pro³ Thi⁵,8 DPhe⁶ CDF⁷-BK)

72. Lys-Lys-Arg-Hyp-Pro-Gly-Phe-Ser-DPhe-Phe-Arg (Lys-Lys-Hyp² DPhe⁷-BK): k(1:1)=0.37; Arg(1.99), Pro(0.96), Gly(0.99), Phe(3.13),Ser(0.94), Hyp(0.98),Lys(2.00)

73. Lys-Lys-Arg-Hyp-Hyp-Gly-Phe-Ser-DPhe-Phe-Arg (Lys-Lys-Hyp²,3 DPhe⁷-Bk): k(1:1)=0.28; Arg(2.03), Gly(0.95), Ser(0.94), Phe(3.08),Lys(2.08), Hyp(1.92)

74. Lys-Lys-Arg-Pro-Pro-Gly-Thi-Ser-DThi-Thi-Arg (Lys-Lys-Thi⁵,8 DThi⁷-BK): k(1:1)=0.22; Arg(2.07), Pro(2.01), Gly(1.04), Ser(0.89),Thi(3.02), Lys(1.96)

75. Lys-Lys-Arg-Pro-Pro-Gly-Thi-Ser-DPal-Thi-Arg (Lys-Lys-Thi⁵,8 DPal⁷-BK): k(1:1)=0.06; Arg(2.12), Pro(1.94), Gly(1.01), Ser(0.87),Thi(1.81), Pal(0.98), Lys(2.22)

76. Lys-Lys-Arg-Pro-Pro-Gly-Thi-DPhe-CDF-Thi-Arg (Lys-Lys-Thi⁵,8 DPhe⁶CDF⁷ -Bk): k(415)=0.11; Arg(2.02), Pro(2.00), Gly(1.02), Phe(1.00),PCF(1.06), Thi(1.85), Lys(2.06)

77. Lys-Lys-Arg-Pro-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Lys-Lys-Hyp³ Thi⁵,8DPhe⁷ -BK): k(1:1)=0.22; Arg(2.05), Pro(0.96), Gly(1.04), Phe(1.03),Ser(0.94), Hyp(1.10), Thi(1.84), Lys(2.04)

78. Lys-Lys-Arg-Hyp-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg (Lys-Lys-Hyp²,3 Thi⁵,8DPhe⁷ -BK): k(1:1)=0.15; Arg(2.00), Pro(2.06), Gly(1.01), Phe(1.03),Ser(0.94), Thi(1.92), Hyp(2.04)

79. Lys-Lys-Arg-Hyp-Pro-Gly-Thi-DPhe-CDF-Thi-Arg (Lys-Lys-Hyp² Thi⁵,8DPhe⁶ CDF⁷ -Bk): k(1:1)=6.69; Arg(1.96), Pro(1.14), Gly(0.96),Phe(1.03), Thi(1.77), PCF(1.06), Lys(1.93), Hyp(1.14)

80. Lys-Lys-Arg-Pro-Hyp-Gly-Thi-DPhe-CDF-Thi-Arg (Lys-Lys-Hyp³ Thi⁵,8DPhe⁶ CDF⁷ -BK): k(1:1)=6.14; Arg(2.04), Pro(0.97), Gly(1.01),Phe(0.98), PCF(1.06), Thi(1.89), Hyp(1.01), Lys(2.05)

81. Lys-Lys-Arg-Hyp-Hyp-Gly-Thi-DPhe-CDF-Thi-Arg (Lys-Lys-Hyp²,3 Thi⁵,8DPhe⁶ CDF⁷ -Bk): k(1:1)=4.88; Arg(2.02), Gly(0.99), Phe(0.96),PCF(1.10), Thi(1.84), Hyp(2.01), Lys(2.06)

82. Arg-Thz-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Thz² Thi⁵,8 DPhe⁷ -BK):k(415)=0.24; Arg(1.97), Pro(1.04), Gly(1.02), Phe(1.06), Thi(1.91)

83. Arg-Pro-Thz-Gly-Thi-Ser-DPhe-Thi-Arg (Thz³ Thi⁵,8 DPhe⁷ -BK):k(1:1)=5.25

84 Arg-Thz-Thz-Gly-Thi-Ser-DPhe-Thi-Arg (Thz²,3 Thi⁵,8 DPhe⁷ -BK):K(1:1)=6.69

85. Arg-Aib-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Aib² Thi⁵,8 DPhe⁷ -BK):k(415)=0.18

86. Arg-Pro-Aib-Gly-Thi-Ser-DPhe-Thi-Arg (Aib³ Thi⁵,8 DPhe⁷ -BK):k(415)=0.24; Arg(2.09), Pro(0.99), Gly(1.05), Ser(0.95), Phe(1.07),Thi(1.94), Aib(0.91)

87. Arg-Aib-Aib-Gly-Thi-Ser-DPhe-Thi-Arg (Aib²,3 Thi⁵,8 DPhe⁷ -BK):k(415)=4.00

88. Arg-Azt-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Azt² Thi⁵,8 DPhe⁷ -BK):k(415)=0.18; Arg(2.07), Pro(0.99), Gly(1.02), Phe(1.04), Ser(0.95),Thi(1.97), Azt(0.99)

89. Arg-Pro-Azt-Gly-Thi-Ser-DPhe-Thi-Arg (Azt³ Thi⁵,8 DPhe⁷ -BK)

90. Arg-Azt-Azt-Gly-Thi-Ser-DPhe-Thi-Arg (Azt²,3 Thi⁵,8 DPhe⁷ -BK)

91. Arg-Inip-Pro-Gly-Thi-Ser-DPhe-Thi-Arg (Inip² Thi⁵,8 DPhe⁷ -BK):k(415)=0.21

92. Arg-Pro-Inip-Gly-Thi-Ser-DPhe-Thi-Arg (Inip³ Thi⁵,8 DPhe⁷ -BK):k(415)=0.18; Arg(2.10), Pro(0.95), Gly(1.04), Phe(1.03), Ser(0.93),Thi(1.95)

93. Arg-Inip-Inip-Gly-Thi-Ser-DPhe-Thi-Arg (Inip²,3 Thi⁵,8 DPhe⁷ -BK):k(415)=0.18

94. Arg-Pro-Pro-Gly-Thi-Ser-DNal-Thi-Arg (Thi⁵,8 DNal⁷ -BK): k(415)=0.32

95. Arg-Pro-Pro-Gly-Thi-Ser-CDF-Thi-Arg (Thi⁵,8 CDF⁷ -BK): k(415)=0.28

96. Arg-Pro-Pro-Gly-Thi-Ser-DTyr-Thi-Arg (Thi⁵,8 DTyr⁷ -BK): k(415)=0.12

97. Arg-Pro-Pro-Gly-Thi-Ser-DVal-Thi-Arg (Thi⁵,8 DVal⁷ -BK): k(415)=0.11

98. Arg-Pro-Pro-Gly-Thi-Ser-DIle-Thi-Arg (Thi⁵,8 DIle⁷ -BK): k(415)=0.16

99. Arg-Pro-Pro-Gly-Thi-Ser-DLeu-Thi-Arg (Thi⁵,8 DLeu⁷ -BK)

100. Arg-Pro-Pro-Gly-Thi-Ser-DTrp-Thi-Arg (Thi⁵,8 DTrp⁷ -BK)

101. Arg-Pro-Pro-Gly-Thi-Ser-DPhg-Thi-Arg (Thi⁵,8 DPhg⁷ -BK)

102. Arg-Pro-Pro-Gly-Thi-Ser-DHis-Thi-Arg (Thi⁵,8 DHis⁷ -BK)

103. Arg-Pro-Pro-Gly-Thi-Ser-DOMT-Thi-Arg (Thi⁵,8 DOMT⁷ -BK):k(415)=0.18

EXAMPLES OF BRADYKININ ANTAGONIST ACTIVITY

The bradykinin antagonists were assayed on isolated rat uterus innatural or induced estrus and on guinea pig ileum, according to thecommonly accepted assay methods for bradykinin and related kinins asdescribed by Trautschold (Handbook of Expt. Pharmacol. Vol 25, SpringerVerlag, pp 53-55, 1970) for inhibition of the myotropic activity ofbradykinin. The inhibition potencies, as determined according to thecommonly accepted manner described by Schild for antagonists ofbiologically active compounds (Br. J. Pharmacol. 2:189,1947), aredetermined on isolated rat uterus (RUT) and isolated guinea pig ileum(GPI). In the assays, a dose-response curve is determined for thereference substance bradykinin. The dose of bradykinin which produced ahalf maximal contraction of tissue is the ED50 dose. An amount ofbradykinin equivalent to twice the ED50 dose is administered to thetissue 30 seconds after the start of incubation of the tissue with adose of antagonist. Doses of antagonist are increased in this protocoluntil pre-incubation with a dose of antagonist reduces the contractionin response to the double ED50 dose of bradykinin to response of asingle ED50 dose of bradykinin. The pA2 value represents the negativelogarithm of the molar concentration of antagonist necessary to reducethe response of a double ED50 dose of bradykinin to that of an ED50dose. One unit of pA2 value represents an order of magnitude change inpotency. For comparison, the negative log of the dose of BK, the dosewhich causes half maximal contraction of the tissues, is commonly knownas the pD2 value. The pD2 value for bradykinin is 7.9 on the rat uterusand 7.4 on the guinea pig ileum.

    ______________________________________                                        POTENCY OF BRADYKININ ANTAGONISTS                                             EX-                                                                           AM-                                                                           PLE                                                                           #    STRUCTURE            pA2/RUT   pA2/GPI                                   ______________________________________                                         1   DPhe.sup.7 --BK                5.0                                        2   DThi.sup.7 --BK                4.6                                        3   DPal.sup.7 --BK      5.0       4.8                                        4   DPhe.sup.6,7 --BK              5.2                                        5   DPhe.sup.6 CDF.sup.7 --BK                                                                          4.9       5.8                                        6   Thi.sup.5,8 DPhe.sup.7 --BK                                                                        6.5       6.3                                        7   Thi.sup.5,8 DThi.sup.7 --BK                                                                        4.2       5.8                                        8   Thi.sup.5,8 DPal.sup.7 --BK                                                                        4.2                                                 10   DArg.sup.0 --DPhe.sup.7 --BK   5.6                                       11   DArg.sup.0 --DPro.sup.3 DPhe.sup.7 --BK                                                            4.0                                                 13   Lys--Lys--DPhe.sup.7 --BK      5.1                                       14   Lys--Lys--Thi.sup.5,8 DPhe.sup.7 --BK                                                              6.0       5.3                                       15   DArg.sup.0 --Thi.sup.5,8 DPhe.sup.7 --BK                                                           5.5       6.1                                       16   DArg.sup.0 --DPro.sup.3 Thi.sup.5,8 DPhe.sup.7 --BK                                                5.2                                                 18   Hyp.sup.2 Thi.sup.5,8 DPhe.sup.7 --BK                                                              5.6                                                 19   Lys--Lys--Hyp.sup.2 Thi.sup.5,8 DPhe.sup.7 --BK                                                    5.8                                                 20   Hyp.sup.3 Thi.sup.5,8 DPhe.sup.7 --BK                                                              7.0       4.7                                       21   DArg.sup.0 -- Hyp.sup.3 Thi.sup.5,8 DPhe.sup.7 --BK                                                7.2                                                 22   Hyp.sup.2,3 Thi.sup.5,8 DPhe.sup.7 --BK                                                            6.7                                                 23   Hyp.sup.2,3 Thi.sup.5,8 DPhe.sup.6 CDF.sup.7 --BK                                                  6.5                                                 64   DArg.sup.0 --Hyp.sup.2 Thi.sup.5,8 DPhe.sup.7 --BK                                                 5.7                                                 65   DArg.sup.0 --Hyp.sup.2,3 Thi.sup.5,8 DPhe.sup.7 --BK                                               7.1                                                 72   Lys--Lys--Hyp.sup.2 DPhe.sup.7 --BK                                                                5.6                                                 77   Lys--Lys--Hyp.sup.3 Thi.sup.5,8 DPhe.sup.7 --BK                                                    6.7                                                 ______________________________________                                    

EXAMPLE OF SPECIFICITY OF KININ ANTAGONISM ON SMOOTH MUSCLE

The specificity of bradykinin antagonists of this invention isdemonstrated by their ability to inhibit the myotropic activity ofbradykinin (BK) and two physiologically important BK-related kinins,kallidin (KAL, Lys-BK) and methionyl-lysyl-BK (MK-BK), but not themyotropic activity induced by non kinin-related peptides, such asangiotensin-II (ANG) or substance-P (SP). In each case, as shown, theBK-related antagonists inhibited contractions produced by BK-relatedagonists, but had no effect on the non-kinin myotropic peptidesubstances. The inhibition potencies are listed as pA2 values asdescribed above.

    __________________________________________________________________________    SPECIFICITY OF BRADYKININ                                                     ANTAGONISTS IN GUINEA PIG ILEUM ASSAY                                                              GUINEA PIG ILEUM                                         EXAMPLE #                                                                             STRUCTURE    BK KAL                                                                              MK-BK                                                                              ANG SP                                        __________________________________________________________________________     1      DPhe.sup.7 --BK                                                                            5.0                                                                              5.6                                                                              6.0  NO  NO                                         6      Thi.sup.5,8 DPhe--BK                                                                       6.3                                                                              6.4                                                                              5.2  NO  NO                                        10      DArg.sup.0 --DPhe.sup.7 --BK                                                               5.6                                                                              6.0                                                                              6.3  NO  NO                                        15      DArg.sup.0 --Thi.sup.5,8 DPhe.sup.7 --BK                                                   6.1                                                                              6.7                                                                              6.4  NO  NO                                        __________________________________________________________________________

EXAMPLE OF THE ANTAGONISM OF BRADYKININ ANTAGONISTS ON RAT BLOODPRESSURE

The in vivo effects of bradykinin antagonists on blood pressure in theanesthetized rat are determined according to the assay described byRoblero, Ryan and Stewart (Res. Commun. Pathol. Pharmacol. 6:207, 1973).When compounds #6 (Thi⁵,8 DPhe⁷ --BK), #10 (DArg⁰ -DPhe⁷ -BK) and #15(DArg⁰ -Thi⁵,8 DPhe⁷ -BK) are infused at a rate of 25 ug/min, theresponse to a 25 mm depressor dose (ED25 mm) of bradykinin is reducedfrom 25 mm to 10 mm. The depressor effect of bradykinin returns to anormal response within 5 minutes of terminating the infusion ofantagonist. The antagonists also produce inhibition of the bradykininresponse when injected as a bolus admixture of bradykinin plusantagonist by either the ia or iv route of administration.

EXAMPLES OF CONFERRING RESISTANCE TO ENZYMATIC DEGRADATION BY EXTENSIONAT THE N-TERMINAL OF BRADYKININ ANALOG

The ability of bradykinin analogs to withstand enzymatic degradation(for example by kininases) in vivo can be conveniently assessed, forexample, by determining residual vasodepressor activity of a particularanalog after a single pass in the pulmonary circulation in theanesthetized rat (J. Roblero, J. W. Ryan, and J. M. Stewart, Res.Commun. Pathol. Pharmacol. 6:207, 1973) following intraaortic andintravenous administration. In this system, using N-terminal substitutedbradykinin analogs (agonists), the following results are obtained:

    ______________________________________                                        RAT PULMONARY DESTRUCTION OF BK ANALOGS                                       MODIFIED AT THE N-TERMINAL                                                    PEPTIDE STRUCTURE                                                                              % DESTRUCTION                                                ______________________________________                                        Bradykinin (BK)  98                                                           Lys--BK          95                                                           DArg--BK         92                                                           DLys--BK         89                                                           Lys--Lys--BK      0                                                           Thi.sup.5,8 --BK 95                                                           Lys--Lys--Thi.sup.5,8 --BK                                                                      0                                                           ______________________________________                                    

The resistance of N-terminal-extended BK analogs to kininasedegradation, especially those with Lys-Lys- extensions, suggests thatlong-acting antagonists of BK activity would be obtained by modifyingthe [D-Phe⁷ ]-BK inhibitors with a Lys-Lys-extension. Additionally,observations that a D-Arg residue added to the N-terminal of BK agoniststends to increase uterine potency without effecting ileum activityprompted the synthesis of D-Arg- extended D-Phe⁷ BK analogs astissue-specific inhibitors. N-terminal extension of D-Phe⁷ BK withLys-Lys- or with the D-Arg residue reduced agonist potency in the uterusassay but has no effect on the antagonism seen in the ileum assay.Similarly, the inhibitory effect of Thi⁵,8 DPhe⁷ BK was diminished onthe uterus with the addition of either Lys-Lys- or D-Arg to theN-terminal. In the ileum assay addition of D-Arg to the very potentThi⁵,8 DPhe⁷ BK antagonist had little effect on inhibitory potency.

EXAMPLES OF CONFERRING TISSUE SELECTIVITY BY MODIFICATION OF BRADYKININANTAGONISTS AT POSITION 2 AND 3

Bradykinin agonists produced by modification of the bradykininnonapeptide sequence exhibit similar potencies in the classic rat uterus(RUT) and guinea pig ileum (GPI) assays relative to bradykinin.Recently, dissociation of smooth muscle activities towards significantlyhigher uterine activity vs ileum activity has been reported inbradykinin analogs containing aminoisobutyric acid (Aib) in position 7(R. J. Vavrek & J. M. Stewart, Peptides 1:231, 1980), and in bradykininanalogs with multiple D-amino acid substitutions in position 6 and 7 (R.J. Vavrek & J. M. Stewart, in Kinins 1984, L.M. Greenbaum, ed, PlenumPress, NY, 1985).

The addition of a D-amino acid residue to the N-terminus of theileum-selective bradykinin antagonist DPhe⁷ -BK (i.e., DArg) does notchange selectivity for inhibiting bradykinin activity on the ileum(i.e., DArg⁰ -DPhe⁷ -BK). Modification of DPhe⁷ -BK at position 3 by thesubstitution of a DPro residue (i.e., DPro³ DPhe⁷ -BK) eliminatesantagonist activity in the ileum assay and destroys agonist activity inthe uterus assay. Replacement of the Pro residue at position 3 of DArg⁰-DPhe⁷ -BK reverses the spectrum of smooth muscle activity towardsuterine- selective antagonism, with complete loss of antagonism ofbradykinin in the ileum assay. Likewise, substitution of the Pro residueat position 3 of the nonspecific antagonist DArg⁰ Thi⁵,8 DPhe⁷ -BK witha DPro residue eliminates antagonism of bradykinin action on the ileum,but full antagonist activity in the uterine assay is retained.

    ______________________________________                                        ANTAGONIST SPECIFICITY IN SMOOTH MUSCLE                                       ASSAYS BY MODIFICATION OF POSITION 3                                          EX-                                                                           AM-                                                                           PLE                                                                           #    STRUCTURE           RUT       GPI                                        ______________________________________                                         1   DPhe.sup.7 --BK     (1% AG)   pA.sub.2 = 5.0                             10   DArg.sup.0 --DPhe.sup.7 --BK                                                                      (0.1% AG) pA.sub.2 = 5.0                             59   DPro.sup.3 --DPhe.sup.7 --BK                                                                      0         0                                          11   DArg.sup.0 --DPro.sup.3 --DPhe.sup.7 --BK                                                         pA.sub.2 = 4.6                                                                          0                                          15   DArg.sup.0 --Thi.sup.5,8 DPhe.sup.7 --BK                                                          pA.sub.2 = 5.2                                                                          pA.sub.2 = 6.1                             16   DArg.sup.0 --DPro.sup.3 Thi.sup.5,8 DPhe.sup.7 --BK                                               pA.sub.2 = 5.2                                                                          0                                          ______________________________________                                         AG = agonist potency relative to BK = 100. pA.sub.2 is the inhibition         potency as defined by Schild (Br. J. Pharmacol. 2:189, 1947).            

Tissue selectivity of bradykinin analogs can also be altered bymodification of the Pro at position 2. It has been found thatsubstitutions of Val, Sar, Pro, Ala, Hyp and Aib(alpha-aminoisobutyrate) at position 2 confer differential tissueselectivity of selected bradykinin analogs in an assay of myotropicactivity determined on guinea pig ileum versus rat uterus (R. J. Vavrekand J. M. Stewart, Peptides, 231-235, 1980). The tissue selectivityconferred upon various bradykinin analogs by such substitutions atposition 2 suggests that similar substitution in any bradykinin analog,including those with antagonist activity, will confer tissueselectivity.

Theraputic applications of the novel bradykinin antagonists include notonly treatmemt for the production of bradykinin or related kinins by theanimal but also the injection of bradykinin related peptides into ananimal as a result of bites and stings. Topical application alone or incombination with subcutaneous utilization of the bradykinin antagonistsof the invention can be employed to treat the effects ofbradykinin-related peptides causing pain, inflammation and swelling.

The therapeutic use of bradykinin antagonists of this invention forother traumatic, inflammatory or pathological conditions which are knownto be mediated by bradykinin or exacerbated by an overproduction ofbradykinin can also be achieved. These conditions include local traumasuch as wounds, burns and rashes, angina, arthritis, asthma, allergies,rhinitis, shock, inflammatory bowel disease, low blood pressure,systemic treatment of pain and inflammation, and low sperm motilitywhich produces male infertility. The present bradykinin antagonists, asdiscussed may be advantageously administered in a variety of waysincluding sublingual absorption as with nitroglycerine or patchadministration using agents for assisting absorption through the skinsuch as for the treatment of angina. Based upon the PA₂ and ED₅₀ datadisclosed in this invention and in the prior art related to agonistpotency, it is possible for one skilled in the art to make adetermination of the dosage of the novel bradykinin antagonists of theinvention.

It is therefore estimated that the dosage range for typical applicationin such conditions as the pain and inflammation of wound, burns andrashes would be 0.1-5 mg/ml; for a nasal spray formulation suitable fortreating rhinitis, allergies and asthma suitable dosage range would be0.1-5 mg/ml; for intravenous formulation suitable for the treatment ofsystemic inflammation, shock, arthritis, allergies, asthma and forincreasing sperm motility, a suitable dosage range would be 0.1-10mg/kg; for an oral formulation for the treatment of inflammatory boweldisease or general pain and inflammation a suitable dosage range wouldbe 10-100 mg/kg. Bradykinin antagonists can also be administeredintravaginally, intrarectally, intrabuccally or any other acceptedinternal application.

As will be recognized by those skilled in the art the present inventionhas a wide range of applicability to providing competitive inhibitors tothe biological activities of bradykinin produced by the body in illness,injury and shock. The advantages of the invention in substituting theL-Pro position 7 with amino acids of the D-configuration to convertbradykinin agonists to antagonists provide a wide variety of specificand competitive antagonists for reducing the known effects ofbradykinin. The additional advantages of the invention of modifying theL-Pro position 7 in conjunction with modifications at the otherpositions of the novel bradykinin antagonists provides a variety ofuseful compounds. It will further be appreciated the present inventionis susceptible to these and other modifications within the parameters ofthe invention without departing from the scope of the following claims.

What is claimed is:
 1. A modified bradykinin type peptide having theformulaA - Arg - B - C - D - W - X - Y - Z - Arg and thepharmaceutically acceptable salts thereof wherein a. A is selected fromthe group comprising(i) hydrogen (ii) Arg (iii) Lys-Lys (iv) Phe (v) Thi(vi) Lys (vii) Met-Lys (viii) Gly-Arg-Met-Lys b. B is selected from thegroup comprising(i) Azetidine-2-Carboxylic Acid (Azt) (ii)Thiazolidine-2-Carboxylic acid (THz) (iii) Isonipecotic acid (Inip) (iv)Pro (v) 2, 3-Dehydroproline (ΔPro) (vi) 4-Hydroxyproline (Hyp) (vii)Hydroxyproline (viii) Aib (ix) Dehydroproline (x) Lys-Lys-Hyp (xi) Ala(xii) Sar (vii) beta-(2-Naphthyl)-Alanine (Nal) (viii) Leu (ix)Lys-Lys-Thi (x) Serine (Ser) (xi) Tyr f. X is selected from the groupcomprising(i) Ser (ii) Gly (iii) Phe (iv) Para-Chloro-D-phenylalanine(CDF) (v) Nal (vi) Pal (vii) Thi (viii) pCl-Phe g. Y is selected fromthe group comprising(i) DNal (ii) DPNF (iii) DPhe (iv) DTyr (v) DPal(vi) DOMT (vii) DThi (viii) DAla (ix) DTrp (x) pNO2 - DPhe (PNF) (xi)DPro (xii) DHis (xiii) D-Homo-Phe (DhPhe) (xiv) pCl-DPhe(CDF) (xv) DPhg(xvi) D-Val (xvii) Lys-Lys-Dtrp (xviii) Lys-Lys-DHis (xix) DIle h. Z isselected from the group comprising(i) Phe (ii) beta-(2-thienyl) alanine(Thi) (iii) 0-methyltyrosine (OMT) (iv) beta-(2-pyridyl) alanine (Pal)(v) para-chloro-L-phenylalanine (CLF) (vi) para-nitrophenylalanine (PNF)(vii) beta-(2-naphthyl) - alanine (Nal) (viii) Leu (ix) Lys-Lys-Thi (x)Tyr
 2. The modified bradykinin type peptide of claim 1 having theformulaDArg-Arg-Hyp-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg
 3. The modifiedbradykinin type peptide of claim 1 having theformulaD-Arg-Arg-Pro-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg
 4. The modifiedbradykinin type peptide of claim 1 having theformulaLys-Lys-Arg-Hyp-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg
 5. The modifiedbradykinin type peptide of claim 1 having theformulaLys-Lys-Arg-Pro-Hyp-Gly-Thi-Ser-DPhe-Thi-Arg
 6. The modifiedbradykinin type peptide of claim 1 having theformulaDArg-Arg-Hyp-Hyp-Gly-Phe-Ser-DPhe-Phe-Arg
 7. The modifiedbradykinin type peptide of claim 1 having theformulaDArg-Arg-Pro-Hyp-Gly-Phe-Ser-DPhe-Phe-Arg
 8. A pharmaceuticalpreparation for treating local pain and inflammation from burns, wounds,cuts, rashes and other such trauma and pathological conditions caused bythe production of bradykinin or related kinins by the animal whichcomprises an effective amount of the modified bradykinin of claim 1sufficient to antagonize bradykinin and a suitable pharmaceuticalcarrier.
 9. A process for treating local pain and inflammation whichcomprises administering an effective amount of the pharmaceuticalpreparation of claim 8 to an animal in need thereof.
 10. Apharmaceutical preparation for treating rhinitis and other such traumaand pathological conditions caused by bradykinin or related kinins inthe animal which comprises an effective amount of the modifiedbradykinin of claim 1 sufficient to antagonize bradykinin and a suitablepharmaceutical carrier.
 11. A process for treating rhinitis whichcomprises administering an effective amount of the pharmaceuticalpreparation of claim 10 to an animal in need thereof.
 12. A modifiedbradykinin type peptide antagonist having the formulaArg - B - C - D -W - X - Y - Z - Arg and the pharmaceutically acceptable salts thereofwherein a. B is selected from the group comprising(i)Azetidine-2-Carboxylic Acid (Azt) (ii) Thiazolidine-2-Carboxylic acid(THz) (iii) Isonipecotic acid (Inip) (iv) Pro (v) 2, 3-Dehydroproline(ΔPro) (vi) 4-Hydroxyproline (Hyp) (vii) Hydroxyproline (viii) Aib (ix)Dehydroproline (x) Lys-Lys-Hyp (xi) Val (xii) Ala (xiii) Sar (xiv) Glyb. C is selected from the group comprising(i) Azetidine-2-Carboxylicacid (Azt) (ii) Thiazolidine-2-Carboxylic acid (THz) (iii) Isonipecoticacid (Inip) (iv) Pro (v) 2, 3-Dehydroproline (ΔPro) (vi)4-Hydroxyproline (Hyp) (vii) Lys-Lys-Hyp (viii) Aib (ix) Val (x) Ala(xi) Sar (xii) Gly c. D is selected from the group comprising(i) Gly(ii) Ala (iii) Sar d. W is selected from the group comprising(i) Phe(ii) beta-(2 Thienyl)-Alanine (THi) (iii) 0-Methyltyrosine (OMT) (iv)beta-(2-Pyridyl) Alanine (Pal) (v) Para-Chloro-L-Phenylalanine (CLF)(vi) Para-nitrophenylalanine (PNF) (vii) beta-(2-Naphthyl)-Alanine (Nal)(viii) Leu (ix) Lys-Lys-Thi (x) Serine (Ser) (xi) Tyr e. X is selectedfrom the group comprising(i) Ser (ii) Gly (iii) Phe (iv)Para-Chloro-D-phenylalanine (CDF) (v) Nal (vi) Pal (vii) Thi f. Y isselected from the group comprising(i) D-aromatic amino acid residue (ii)substituted D-aromatic amino acid residue g. Z is selected from thegroup comprising(i) Phe (ii) beta-(2-thienyl) alanine (Thi) (iii)0-methyltyrosine (OMT) (iv) beta-(2-pyridyl) alanine (Pal) (v)para-chloro-L-phenylalanine (CLF) (vi) para-nitrophenylalanine (PNF)(vii) beta-(2-naphthyl) - alanine (Nal) (viii) Leu (ix) Lys-Lys-Thi (x)Tyr
 13. A pharmaceutical preparation for treating local pain andinflammation from burns, wounds, cuts, rashes and other such trauma andpathological conditions caused by the production of bradykinin orrelated kinins by the animal which comprises an effective amount of themodified bradykinin of claim 12 sufficient to antagonize bradykinin anda suitable pharmaceutical carrier.
 14. A process for treating local painand inflammation which comprises administering an effective amount ofthe pharmaceutical preparation of claim 13 to an animal in need thereof.15. A pharmaceutical preparation for treating rhinitis and other suchtrauma and pathological conditions caused by bradykinin or relatedkinins in the animal which comprises an effective amount of theantagonist bradykinin of claim 12 sufficient to antagonize bradykininand a suitable pharmaceutical carrier.
 16. A process for treatingrhinitis which comprises administering to a host the pharmaceuticalpreparation of claim 15 to an animal in need thereof.
 17. A modifiedbradykinin type peptide antagonist having the formulaArg- Pro - Pro -Gly - Phe - Ser - Y - Phe - Arg and pharmaceutically acceptable saltsthereof wherein Y is DPhe, beta-2-thienyl-DAla (DThi),beta-2-pyridyl-DAla (DPal), beta-2-naphthyl-DAla(DNal), DHis,D-homo-Phe(DhPhe), 0-methyl-DTyr(DOMT), D-alphaphenyl-Gly(DPhg), DTrp,DTyr, pNO₂ -DPhe(PNF) or pCl-DPhe(CDF).
 18. A pharmaceutical preparationfor treating local pain and inflammation from burns, wounds, cuts,rashes and other such trauma and pathological conditions caused by theproduction of bradykinin or related kinins by the animal which comprisesan effective amount of the modified bradykinin of claim 17 sufficient toantagonize bradykinin and a suitable pharmaceutical carrier.
 19. Aprocess for treating local pain and inflammation which comprisesadministering an effective amount of the pharmaceutical preparation ofclaim 18 to an animal in need thereof.
 20. A pharmaceutical preparationfor treating rhinitis and other such trauma and pathological conditionscaused by bradykinin or related kinins in the animal which comprises aneffective amount of the antagonist bradykinin of claim 18 sufficient tomodified bradykinin and a suitable pharmaceutical carrier.
 21. A processfor treating rhinitis which comprises administering an effective amountof the pharmaceutical preparation of claim 19 to an animal in needthereof.